Hochdurchsatz-Analyse pathogener Eigenschaften Herpesvirus-8-kodierter Gene in Endothelzellen

卡波西肉瘤（KS）是一种血管增生性疾病。人类疱疹病毒-8 （HHV-8）在KS中具有病原学作用。目前尚不清楚HHV-8的哪些基因可能引发KS的发展。在这个项目中，我们将系统地确定HHV-8编码的84个基因在原代人内皮细胞培养中的作用。特别是与肿瘤相关的功能，影响细胞增殖、单核细胞粘附、迁移和凋亡将被研究。此外，每个基因的作用将与其他HHV-8基因以及原癌基因c-myc和ras结合进行测试。最后，整个方法将在受到炎症细胞因子、血管生成生长因子和HIV-1-Tat蛋白不同外部刺激的内皮细胞中进行。为了实现这一目标，将建立一种高通量的表达质粒反向转染系统，可以并行确定数百个基因的功能。本研究将鉴定在内皮细胞中具有最高致瘤激活潜能的HHV-8基因。此外，我们的方法可以提供一个有用的平台技术来确定任何一组病毒和细胞基因在Schwerpunktprogramm范围内的血管致病功能。

Kaposi's sarcoma (KS) is an angioproliferative disease. Human herpesvirus-8 (HHV-8) has an etiologic role in KS. It is unknown, as yet, which genes of HHV-8 may trigger KS development. In this project we will systematically determine the effects of each of the 84 genes encoded by HHV-8 in primary human endothelial cell cultures. Particularly tumor relevant functions, affecting cell proliferation, adhesiveness for monocytes, migration and apoptosis will be investigated. In addition, the effect of each gene will be tested in combination with every other HHV-8 gene and with the proto-oncogenes c-myc and ras. Finally, the whole approach will be performed with endothelial cells that have been subjected to different external stimulations by inflammatory cytokines, angiogenic growth factors and the HIV-1-Tat protein. To this goal, reverse transfection of arrayed expression plasmids will be established as a high throughput transfection system which allows to determine the functions of hundreds of genes in parallel. This study will identify the genes of HHV-8 with the highest tumorigenic activation potential in endothelial cells. Moreover, our approach may provide a useful platform technology to determine the vasculopathogenic function of any set of viral and cellular genes in the scope of the Schwerpunktprogramm.